A vascularized human lung-on-a-chip reveals compartmentalized inflammatory responses in bacterial pneumonia

A vascularized human lung-on-a-chip reveals compartmentalized inflammatory responses in bacterial pneumonia | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Article A vascularized human lung-on-a-chip reveals compartmentalized inflammatory responses in bacterial pneumonia Kelsie Volek, Hellen Kang, Nicole Rosin, Mohammad Rehan, Theodore Nelson, and 6 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-3343362/v1 This work is licensed under a CC BY 4.0 License Status: Under Review Version 1 posted You are reading this latest preprint version Abstract The lung serves as a primary organ for gas exchange in the human body. However, during clinically apparent lung insults such as bacterial pneumonia, the blood-alveolar barrier is disrupted, immune cells are recruited, and lung biomechanics are altered. These processes impair lung function acutely with major contributions from the lung vasculature. The pulmonary vasculature normally provides an 8km long vascular network interfacing with the 50-100m2 epithelial surface of the lung. This vasculature is a critical regulator of lung development, homeostasis, and function; yet to date, 3D organ-on-a-chip lung models have largely relied on planar vascular structures that do not recapitulate a true microvascular compartment. Here, we generate a human vascularized lung on a chip (LoC) containing a ventilated epithelial lined air-liquid interface surrounded by human lung fibroblasts and a functional microvascular network. We perform detailed transcriptomic, morphologic, and functional characterization of the LoC to demonstrate the physiologic relevance of this model and examine acute inflammation during bacterial pneumonia. Using scRNAseq we observe enhanced cellular differentiation, function, and crosstalk in LoC 3D co-culture. During bacterial infection with Streptococcus pneumoniae, the most common bacterial cause of human pneumonia, we identify a sequence of events where infection causes compartmentalized inflammatory responses and intravascular immune cell recruitment. This precedes complete air-blood barrier breakdown, tissue destruction associated with necroptotic cell death, and loss of tissue elasticity. Finally, using selective protease inhibitors we demonstrate a role for ADAM10 sheddase in driving neutrophil infiltration and epithelial dysfunction during pneumococcal pneumonia. This model provides a novel platform to mechanistically explore lung homeostasis and human lung diseases with potential for integration into existing human drug development pipelines. Physical sciences/Engineering/Biomedical engineering Biological sciences/Biotechnology/Tissue engineering Lung Organ on Chip Microphysiological Systems Pneumonia Inflammation Full Text Additional Declarations There is NO Competing Interest. Supplementary Files TableS1EndothelialOrganSpecificGenes.xlsx bulk RNAseq of HLMEC, HUVEC, HBMEC, HLSEC TableS2LoCSuerattop20markers.csv LoC scRNAseq Seurat Clusters TableS3LoCEndothelial.csv LoC Endothelial scRNAseq DEGs TableS4LoCEpithelial.csv LoC Epithelial scRNAseq DEGs TableS5LoCFibroblast.csv LoC Fibroblast scRNAseq DEGs TableS6LoC3Dconnectome.xlsx Table S6 – LoC scRNAseq Connectomics MovieS1.mp4 LoC Ventilation MovieS2.mp4 Streptococcus pneumoniae treated LoC exhibit tissue failure during mechanical ventilation RSMALJD1800285toreviewMRGFINALDec132023.pdf Reporting Summary Cite Share Download PDF Status: Under Review Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. 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Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-3343362","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":352083090,"identity":"eee92e08-ae50-420c-b35f-96b8f145be03","order_by":0,"name":"Kelsie Volek","email":"","orcid":"","institution":"University of Calgary","correspondingAuthor":false,"prefix":"","firstName":"Kelsie","middleName":"","lastName":"Volek","suffix":""},{"id":352083091,"identity":"6db431a5-5f3f-4a06-bab5-14bba9dfb4dd","order_by":1,"name":"Hellen Kang","email":"","orcid":"","institution":"University of 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